中华骨质疏松和骨矿盐疾病杂志
中華骨質疏鬆和骨礦鹽疾病雜誌
중화골질소송화골광염질병잡지
CHINESE JOURNAL OF OSTEOPOROSIS AND BONE MINERAL RESEARCH
2014年
1期
42-47
,共6页
唐雯菁%杜艳萍%洪维%李慧林%程群
唐雯菁%杜豔萍%洪維%李慧林%程群
당문정%두염평%홍유%리혜림%정군
骨髓间充质干细胞%甲状旁腺素%增殖%分化%旁分泌
骨髓間充質榦細胞%甲狀徬腺素%增殖%分化%徬分泌
골수간충질간세포%갑상방선소%증식%분화%방분비
mesenchymal stem cell%parathyroid hormone%proliferation%differentiation%paracrine
目的：观察甲状旁腺素（PTH）对体外培养人骨髓间充质干细胞（hMSCs）增殖、分化、干性（ stemness ）和旁分泌功能的作用。方法采用标准培养基体外培养人骨髓间充质干细胞，并将其分为对照组和PTH干预组，用细胞计数测试和BrdU增殖检测细胞增殖变化。用碱性磷酸酶染色、油红染色、阿利新蓝染色检测细胞分化情况。用Real-time PCR检测hMSCs OCT-4、Nanog、 SOX-2、 CCND1、 C-MYC等基因表达情况。用ELISA方法检测hMSCs培养上清液基质细胞衍生因子-1（SDF-1）、胰岛素样生长因子（IGF-1）、血管内皮生长因子（ VEGF）、骨形态发生蛋白2（ BMP2）和Angiopoietin1蛋白水平。结果使用10-8 mol/L～10-9 mol/L的PTH干预6 d可明显促进hMSCs增殖（ P＜0.05）， PTH 10-8 mol/L促进成骨、软骨分化，抑制脂肪分化。PTH 10-8 mol/L可明显诱导CCND1和C-MYC基因表达（均P＜0.01），抑制OCT-4表达（ P＜0.01），抑制Nanog和SOX-2等表达（均P＜0.05）），升高培养液中SDF-1、 IGF-1、 VEGF、 BMP2和血管生成素1（ Angio-poietin 1）蛋白水平（均P＜0.05）。结论 PTH能促进hMSCs增殖，调节其分化，但不能维持hMSCs的干性。调节hMSCs的旁分泌功能对局部微环境合成代谢、细胞迁移和血管发生产生影响。
目的：觀察甲狀徬腺素（PTH）對體外培養人骨髓間充質榦細胞（hMSCs）增殖、分化、榦性（ stemness ）和徬分泌功能的作用。方法採用標準培養基體外培養人骨髓間充質榦細胞，併將其分為對照組和PTH榦預組，用細胞計數測試和BrdU增殖檢測細胞增殖變化。用堿性燐痠酶染色、油紅染色、阿利新藍染色檢測細胞分化情況。用Real-time PCR檢測hMSCs OCT-4、Nanog、 SOX-2、 CCND1、 C-MYC等基因錶達情況。用ELISA方法檢測hMSCs培養上清液基質細胞衍生因子-1（SDF-1）、胰島素樣生長因子（IGF-1）、血管內皮生長因子（ VEGF）、骨形態髮生蛋白2（ BMP2）和Angiopoietin1蛋白水平。結果使用10-8 mol/L～10-9 mol/L的PTH榦預6 d可明顯促進hMSCs增殖（ P＜0.05）， PTH 10-8 mol/L促進成骨、軟骨分化，抑製脂肪分化。PTH 10-8 mol/L可明顯誘導CCND1和C-MYC基因錶達（均P＜0.01），抑製OCT-4錶達（ P＜0.01），抑製Nanog和SOX-2等錶達（均P＜0.05）），升高培養液中SDF-1、 IGF-1、 VEGF、 BMP2和血管生成素1（ Angio-poietin 1）蛋白水平（均P＜0.05）。結論 PTH能促進hMSCs增殖，調節其分化，但不能維持hMSCs的榦性。調節hMSCs的徬分泌功能對跼部微環境閤成代謝、細胞遷移和血管髮生產生影響。
목적：관찰갑상방선소（PTH）대체외배양인골수간충질간세포（hMSCs）증식、분화、간성（ stemness ）화방분비공능적작용。방법채용표준배양기체외배양인골수간충질간세포，병장기분위대조조화PTH간예조，용세포계수측시화BrdU증식검측세포증식변화。용감성린산매염색、유홍염색、아리신람염색검측세포분화정황。용Real-time PCR검측hMSCs OCT-4、Nanog、 SOX-2、 CCND1、 C-MYC등기인표체정황。용ELISA방법검측hMSCs배양상청액기질세포연생인자-1（SDF-1）、이도소양생장인자（IGF-1）、혈관내피생장인자（ VEGF）、골형태발생단백2（ BMP2）화Angiopoietin1단백수평。결과사용10-8 mol/L～10-9 mol/L적PTH간예6 d가명현촉진hMSCs증식（ P＜0.05）， PTH 10-8 mol/L촉진성골、연골분화，억제지방분화。PTH 10-8 mol/L가명현유도CCND1화C-MYC기인표체（균P＜0.01），억제OCT-4표체（ P＜0.01），억제Nanog화SOX-2등표체（균P＜0.05）），승고배양액중SDF-1、 IGF-1、 VEGF、 BMP2화혈관생성소1（ Angio-poietin 1）단백수평（균P＜0.05）。결론 PTH능촉진hMSCs증식，조절기분화，단불능유지hMSCs적간성。조절hMSCs적방분비공능대국부미배경합성대사、세포천이화혈관발생산생영향。
Objective To investigate the effects of parathyroid hormone (PTH) on proliferation, differentia-tion, stemness and paracrine of human mesenchymal stem cells ( hMSCs ) .Methods hMSCs were bought from Lonza . hMSCs were divided into two groups:CON, hMSCs were cultured by normal medium;PTH, cultured by normal medium with PTH.Cell proliferation was detected by cell counting and BrdU incorporation .Cell differentiation was detected using ALP, Oil Red O and Alcian Blue staining .mRNA expression of OCT-4, Nanog, SOX-2 and CCND1, C-MYC of hMSCs were measured by Realtime PCR .Protein concentrations of SDF-1, IGF-1, VEGF, BMP2 and Angiopoietin1 in hMSCs culture medium were tested by ELISA .Results PTH 10 -8 -10 -9 mol/L increased proliferation of hMSCs at day 6 ( P<0.05 ) , and PTH 10 -8 mol/L promoted osteogenesis and chondrogenesis , inhibited adipogenesis of hMSCs .PTH 10 -8 mol/L induce the gene expression of CCND 1 ( P<0.01 ) and C-MYC ( P<0.01 ) , and suppressed the gene expression of OCT-4 (P<0.01), Nanog (P<0.05) and SOX-2 (P<0.05) in vitro.PTH 10 -8 mol/Lalso increased the protein concentration of SDF-1 (P<0.05), IGF-1 (P<0.05), BMP2 (P<0.05) and Angiopoietin1 (P<0.05) in cell cul-ture medium which shows great important in regulation and communication of hMSCs with other tissue around microenvi -ronment .Conclusion PTH greatly regulated hMSCs differentiation , and moderately promoted hMSCs proliferation , how-ever, PTH cann't maintain hMSCs stemness .PTH regulated paracrine action of hMSCs , which promoted anabolism , mi-gration and angiogenesis .
